BPN794: Bubble-Free Microfluidic PCR

Project ID

BPN794

Website

Start Date

Thu 2015-Feb-05 01:38:31

Last Updated

Tue 2015-Aug-25 10:37:57

Abstract

Microfluidic polymerase chain reaction (PCR) has been of great interest owing to its
ability to perform rapid and specific nucleic acid amplification and analysis on small
volumes of samples. One of the major drawbacks of microfluidic PCR is bubble generation
and reagent evaporation, which can cause malfunctions. Here, we propose a bubble-free
microfluidic PCR device via controlled fluid transfer. We design the PCR chamber
surrounding circumferential chamber allows a guided-fluid transport of generated bubble to
circumferential chamber through thin nanoporous PDMS sidewall. We accomplish the
theoretical modeling of formation bubble behavior and removal of bubbles. We solve the
generation of bubbles by inhibiting mass transport along a vertical direction via a thin
impermeable polyethylene (PE) as a top layer, and the fast gas transport through the
circumferential ring barrier of nanoporous poly(dimethylsiloxane). We demonstrate stand-
alone self-powered sample loading and the guided-fluid transport of bubble escapement
during thermal cycling, and accomplish the successful amplification of the cMET gene, a
nucleic acid (NA) biomarker for lung cancer. In addition, an ultrafast digital PCR test
on chip was completed in less than 3 min using a high powered Peltier-based thermal cycler
without bubbles. This approach will result in a new paradigm for ultrafast molecular
diagnosis on chip and can facilitate NA-based nearly instantaneous diagnostics for point-
of-care testing and for personalized and preventive medicine.